Substances for reducing occurrence of major cardiac events comprising red yeast rice extract and omega-3 polyunsaturated fatty acid or derivative thereof

ABSTRACT

Medicaments and therapeutic compositions comprise Red Yeast Rice extract and omega-3 polyunsaturated fatty acids and/or derivatives thereof, e.g., DHA, derivatives of DHA, EPA, derivatives of EPA or mixtures thereof. One source of the fatty acids or derivatives thereof is fish oil. The compositions are useful for lowering cholesterol and/or triglyceride levels in a subject.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation of U.S. patent application Ser. No.12/630,686, filed Dec. 3, 2009, which is a continuation-in-part of U.S.patent application Ser. No. 11/757,340 filed Jun. 1, 2007, the entirecontents of which are incorporated herein by reference. Allpublications, patents, patent applications, databases and otherreferences cited in this application, all related applicationsreferenced herein, and all references cited therein are incorporatedherein by reference in their entirety as if restated here in full and asif each individual publication, patent, patent application, database orother reference were specifically and individually indicated to beincorporated by reference.

BACKGROUND OF THE INVENTION Field of the Invention

The invention relates to compositions and the use of such compositionsin nutritional supplements and medicaments, wherein the compositions arecombinations of Red Yeast Rice extract, having hydroxymethylglutaryl-CoA(HMG-CoA) reductase inhibitor activity, with omega-3 polyunsaturatedfatty acid or derivatives thereof derived from fish oils or othersources of omega-3 polyunsaturated fatty acids or derivatives thereof.

Description of the Prior Art

Statins (which are members of a group of HMG-CoA reductase inhibitors)are a group of hypolipidemic agents, used as pharmaceutical agents tolower cholesterol levels in people with or at risk for cardiovasculardisease. Statins lower cholesterol by inhibiting the enzyme HMG-CoAreductase, which is the rate-limiting enzyme of the mevalonate pathwayof cholesterol synthesis. Inhibition of this enzyme in the liverstimulates low-density lipoprotein (LDL) receptors, resulting in anincreased clearance of LDL, so-called “bad cholesterol,” from thebloodstream and a decrease in blood cholesterol levels.

Statins are potent cholesterol-lowering agents, and have been reportedto lower LDL cholesterol by 30-50% (see Jones, P., Kafonek, S., Laurora,I., et al., “Comparative dose efficacy study of atorvastatin versussimvastatin, pravastatin, lovastatin, and fluvastatin in patients withhypercholesterolemia (the CURVES study),” Am J Cardiol 1998; 81-(5);582-7). Statins are classified as either synthetic or fermentationderived. Lovastatin was isolated from a strain of Aspergillus terreusand it was the first statin approved by the FDA as a drug (August 1987).Lovastatin is a water insoluble, white crystalline solid. The aqueousinsolubility of lovastatin leads to inadequate dissolution ingastrointestinal fluids and, hence, poor absorption, distribution, andtargeted organ delivery. Solubility of lovastatin is enhanced byreaction with 3-cyclodextrin, an oligosaccharide which improves thesolubility of lovastatin. The improvement of aqueous solubility in sucha case is a valuable goal to improve therapeutic efficacy. Lovastatincan also produce slight to moderate increases in high densitylipoproteins (HDL) (10-20%), and slight decreases in triglycerides(5-10%). The usual daily dose of lovastatin is 20-80 mg/day. The statindrugs include lovastatin, pravastatin, fluvastatin, atorvastatin,simvastatin, rosuvastatin, and cerivastatin.

Compounds similar to lovastatin have also been found in a naturalfermentation product known as Red Yeast Rice. These compounds are alsoHMGCoA reductase inhibitors. A monograph published in AlternativeMedicine Review (Volume 9, Number 2, 2004) reports that the HMG-CoAreductase inhibitor activity in Red Yeast Rice comes from a naturallyoccurring family of nine compounds called “monacolins,” each of whichhas HMG-CoA reductase inhibitor activity. Additional active ingredientsin Red Yeast Rice include sterols (beta-sitosterol, campesterol,sigmasterol, and sapogenin), isoflavones, and monounsaturated fattyacids (see Heber D, et al., Cholesterol lowering effects of proprietaryChinese red yeast rice dietary supplement, Am J Clin Nutr 1999,69:231-236). One of the monacolins in Red Yeast Rice, monacolin K, issaid to be the lactone form of the statin drug lovastatin, which isconverted to the active acid form in vivo by the liver.

Red Yeast Rice is a common foodstuff in Asian countries where theaverage daily intake is 14-55 grams. The nutritional supplement derivedfrom Red Yeast Rice is known as Red Yeast Rice extract. It is obtainedby drying the fermented product of rice on which the yeast Monascuspupureus has been grown and extracting the dried product with a solvent,usually aqueous ethanol or water. The Red Yeast Rice extract thusproduced typically contains about 0.2 wt. % monacolin K and about 0.5wt. % total monacolins.

U.S. Pat. No. 6,046,022, issued Apr. 4, 2000 to Zhang, et al (PekingUniversity), discloses some methods of making high lovastatin (monacolinK) Red Yeast Rice and using Red Yeast Rice and Red Yeast Rice extract.U.S. Pat. No. 6,046,022 is hereby incorporated by reference herein inits entirety.

U.S. Pat. Nos. 6,541,005; 6,436,406; 6,495,173; 6,544,525; 6,576,242;and 6,541,006, issued Apr. 1, 2003; Aug. 20, 2002; Dec. 17, 2002; Apr.8, 2003; Jun. 10, 2003; Apr. 1, 2003; and Jun. 25, 2002, respectively toYegorova and U.S. Pat. No. 6,410,521, issued Jun. 25, 2002 to Mundy etal., disclose methods of using Red Yeast Rice. All of these patents areincorporated herein by reference in their entirety.

U.S. Patent Application Publication No. 20060211763, published Sep. 21,2006 by Fawzy et al., discloses a statin drug dissolved in a solventsystem comprising natural or synthetic omega-3 fatty acids and U.S.Patent Application Publication No. 20060034815, published Feb. 16, 2006by Guzman et al., discloses omega-3 oil solutions of one or morestatins.

Omega-3 polyunsaturated fatty acids and derivatives thereof can bederived from fish oils and are known to reduce serum triglycerides (seeAbe Y, El-Masri B, et al, Soluble cell adhesion molecules inhypertriglyceridemia and potential significance on monocyte adhesion.Arteriosler Thromb Vasc Biology 1998:18:723-731) and adverse coronaryevents. The principal active ingredients in fish oil are5,8,11,14,17-eicosapentaenoic acid (eicosapentaenoic acid (EPA), 20:5(n-3)) and 4,7,10,13,16,19-docosahexaenoic acid (docosahexaenoic acid(DHA), 22:6 (n-3)). EPA and DHA have been given at a combined dose of 4g/day for seven months to hypertriglyceridemic patients resulting in areduction of 47% in triglycerides (see Ridker, Paul, Effects of n-3Fatty Acid Therapy on Lipids and sCAMs—inflammatory Markers,Pharmacotherapy and Clinical Trials, Lipids Online.org, posted: Oct. 3,2001, reviewed Oct. 4, 2001).

The effects of statin drugs and omega-3 polyunsaturated fatty acids orderivatives thereof have been reported to be cumulative. When 59patients who were already receiving 10-40 mg daily of the statinsimvastatin were given 2 grams twice daily of EPA+DHA, there was afurther sustained significant decrease of 20-30% in triglycerides (seeDurington P, Bhatnager, et al, An omega-3 polyunsaturated fatty acidconcentration administered for one year decreased triglycerides insimvastin treated patients with CM. Heart 2001:85(5) 544-548).

Omega-3 polyunsaturated fatty acids and derivatives thereof are alsowell known to those skilled in the art to reduce inflammation, decreasearrhythmias, decrease risk of sudden cardiac death and cardiac arrest.

SUMMARY OF THE INVENTION

Further provided in accordance with the present invention is atherapeutic composition comprising (1) at least one omega-3polyunsaturated fatty acid, at least one pharmaceutically acceptableomega-3 polyunsaturated fatty acid derivative or mixtures thereof, and(2) Red Yeast Rice extract.

The present invention further provides such a therapeutic compositionwherein component (1) comprises EPA, derivatives of EPA, DHA,derivatives of DHA or mixtures thereof. The therapeutic compositions maybe compositions wherein the derivatives of EPA and derivatives of DHAare glycerides. Further provided are compositions wherein component (1)is a mixture comprising about 35 wt. % triglycerides of EPA and about 25wt. % triglycerides of DHA.

The present invention further provides a therapeutic composition whereincomponent (1) is a mixture comprising at least about 60 wt. % of acombination of EPA and DHA in a weight ratio of EPA:DHA of from about1.4:1 to about 5:1, wherein the combination is at least about 60% in thetriglyceride form of the EPA and DHA and the balance is at least about80% mono- and di-glycerides. Also provided are compositions wherein thecombination comprises about 65 wt. % triglycerides of EPA and about 15wt. % triglycerides of DHA or wherein the combination comprises about 75wt. % triglycerides of EPA and about 15 wt. % triglycerides of DHA. Alsoprovided are compositions wherein the combination is at least about 80%in the triglyceride form, at least about 90% in the triglyceride form,at least about 98% in the triglyceride form, or least about 98% in thetriglyceride form and the remainder is monoglycerides, diglycerides orboth. The present invention further provides therapeutic compositionswherein the combination comprises about 65 wt. % triglycerides of EPAand about 15 wt. % triglycerides of DHA.

In accordance with the present invention there is further providedcompositions wherein the Red Yeast Rice extract comprises at least about0.1 wt. % monacolin K, wherein Red Yeast Rice extract comprises at leastabout 0.2 wt. % monacolin K and at least about 0.4 wt. % totalmonacolins, and wherein the Red Yeast Rice extract comprises at leastabout 0.4 wt. % monacolin K.

The present invention also provides a dose of the medicament ortherapeutic composition wherein the dose of medicament or therapeuticcomposition comprises about 1000 mg of EPA and/or derivatives of EPAplus DHA and/or derivatives of DHA and about 1200 mg of Red Yeast Riceextract, wherein the dose comprises about 2000 mg of EPA and/orderivatives of EPA plus DHA and/or derivatives of DHA and about 2400 mgof Red Yeast Rice extract, or wherein the dose comprises about 4000 mgof EPA and/or derivatives of EPA plus DHA and/or derivatives of DHA andabout 2400 mg of Red Yeast Rice extract

In accordance with the present invention there is further provided amethod of reducing serum cholesterol, triglycerides or both in a subjectcomprising administering to the subject a dosage comprising theabove-recited therapeutic compositions.

DETAILED DESCRIPTION OF EMBODIMENTS

The present invention relates to a medicament comprising Red Yeast Riceextract dispersed in fish oil. In some embodiments, the medicamentincludes compositions in which the fish oil comprises at least about 60%of omega-3 oils, or at least about 70% omega-3 oils. In someembodiments, the medicament includes compositions in which the omega-3oils comprise about 50% EPA and about 35% DHA, or in which the omega-3oils comprise about 69% EPA and about 16% DHA. In some embodiments, themedicament includes compositions in which the Red Yeast Rice extractcomprises at least about 0.1 wt. % monacolin K. In some embodiments, themedicament includes compositions in which the Red Yeast Rice extractcomprises at least about 0.2 wt. % monacolin K and at least about 0.4wt. % total monacolins. In some embodiments, the weight ratio of fishoil to Red Yeast Rice extract is in the range between 1.4 and 2.8. Insome embodiments, the medicament further comprises a dispersing agent.In some embodiments, the dispersing agent comprises 3% lysine and 2%bamboo. In some embodiments, the medicament further comprises a softgelatin capsule into which the fish oil, Red Yeast Rice extract anddispersing agent are loaded. In some embodiments, the medicamentcomprises a daily dose of the medicament which is delivered by anintegral number of capsules. In some embodiments, the daily dose ofmedicament comprises about 1000 mg of EPA and/or derivative of EPA plusDHA and/or derivative of DHA and about 1200 mg of Red Yeast Riceextract, or the daily dose of medicament comprises about 2000 mg of EPAand/or derivative of EPA plus DHA and/or derivative of DHA and about1200 mg of Red Yeast Rice extract, or the daily dose of medicamentcomprises about 2000 mg of EPA and/or derivative of EPA plus DHA and/orderivative of DHA and about 2400 mg of Red Yeast Rice extract, or thedaily dose of medicament comprises about 4000 mg of EPA and/orderivative of EPA plus DHA and/or derivative of DHA and about 2400 mg ofRed Yeast Rice extract. In some embodiments, the medicament furthercomprises an antioxidant. In some embodiments, the antioxidant is chosenfrom the group consisting of rosemary, vitamin E, astaxanthine,carnitine, ascorbyl palmitate, and tocopherols.

The present invention further relates to a method of reducing serumcholesterol, triglycerides or both in a subject comprising administeringa daily dosage comprising EPA and/or derivative of EPA, DHA and/orderivative of DHA, and Red Yeast Rice extract. In some embodiments, thedaily dosage comprises about 4000 mg of EPA and/or derivative of EPAplus DHA and/or derivative of DHA and about 2400 mg of Red Yeast Riceextract, or the daily dosage comprises about 1000 mg of EPA and/orderivative of EPA plus DHA and/or derivative of DHA and about 1200 mg ofRed Yeast Rice extract, or the daily dosage comprises about 2000 mg ofEPA and/or derivative of EPA plus DHA and/or derivative of DHA and about1200 mg of Red Yeast Rice extract, or the daily dosage comprises about2000 mg of EPA and/or derivative of EPA plus DHA and/or derivative ofDHA and about 2400 mg of Red Yeast Rice extract.

The invention also relates to compositions comprising Red Yeast Riceextract dispersed in omega-3 polyunsaturated fatty acids or derivativesthereof, and the use of such compositions to treat a subject. The RedYeast Rice extract preferably comprises at least one and preferably morethan one monacolin compound. In some embodiments, the monacolin compoundcomprises monacolin K. In some embodiments, the monacolin compoundscomprise monacolin K and at least one other monacolin compound. In someembodiments, the monacolin compounds comprise all of the monacolincompounds in Red Yeast Rice. In some embodiments, the Red Yeast Riceextract contains about 0.2 wt. % monacolin K and about 0.5 wt. % oftotal monacolins.

The compositions of the present provide several advantages over the useof lovastatin to reduce cholesterol and triglyceride level in a subject.For instance, the Red Yeast Rice extract is water soluble, whereaslovastatin is not. As noted above, the water insolubility of lovastatinleads to inadequate dissolution in gastrointestinal fluids and, hence,poor absorption, distribution, and targeted organ delivery. While thewater solubility of lovastatin can be enhanced, it is believed that thewater soluble Red Yeast Rice extract will enter the subjects systemeasier than lovastatin.

It is also emphasized that Red Yeast Rice extract can produce betterlipid reducing results at lower dosages (based on the amount ofmonacolin in the Red Yeast Rice extract) than lovastatin. This reducesthe risk of undesirable and possibly harmful side effects in thesubject.

The Red Yeast Rice extract is prepared by fermenting white rice,preferably non-glutinous white rice, with Monascus purpureus strain ofyeast by culturing said Monascus purpureus strain in a culture mediumcomprising rice at a temperature of about 15° C. to about 35° C. for aperiod of about 2 to about 20 days to provide a crude fermentationproduct containing Red Yeast Rice; drying said crude fermentationproduct to obtain Red Yeast Rice, extracting said Red Yeast Rice with asolvent to provide an extract; and drying said extract to remove thesolvent and produce Red Yeast Rice extract. The solvent is preferablyeither aqueous ethanol or water. Other culture media may also be addedto the rice. For example, sugar; an additional carbon source chosen fromthe group consisting of glycerine, malt, and potato juice; and thickbeet juice or mixtures thereof may be used. In addition, a defoamer maybe added.

In some embodiments, the Red Yeast Rice extract used in the compositionsof the present invention contains at least about 0.1 wt. % monacolin K.In some embodiments, the Red Yeast Rice extract contains at least about0.15 wt. %, at least about 0.2 wt. %, at least about 0.25 wt. %, atleast about 0.3 wt. %, at least about 0.35 wt. % or at least about 0.4wt. % monacolin K. In some embodiments, the Red Yeast Rice extractcontains at least about 0.4 wt. %, at least about 0.45 wt. % or at leastabout 0.5 wt. % of total monacolins. In some embodiments, the Red YeastRice extract used in the compositions of the present invention containsat least about 0.4 wt. % monacolin K.

Red Yeast Rice extracts are readily available in commerce in the UnitedStates and may be purchased already prepared. Preferred typical dailydose of Red Yeast Rice extract is about 1.2 to about 2.4 grams per day,which were the dosages used in human clinical trials.

Omega-3 Polyunsaturated Fatty Acids

As used herein, the term “omega-3 polyunsaturated fatty acid(s)” refersto a family of unsaturated fatty carboxylic acids that have in common acarbon-carbon bond in the n-3 position (i.e., the third bond from themethyl end of the molecule). Typically, they contain from about 16 toabout 24 carbon atoms and from three to six carbon-carbon double bonds.Omega-3 polyunsaturated fatty acids can be found in nature, and thesenatural omega-3 polyunsaturated fatty acids frequently have all of theircarbon-carbon double bonds in the cis-configuration.

Examples of omega-3 polyunsaturated fatty acids include, but are notlimited to, 7,10,13-hexadecatrienoic acid (sometimes abbreviated as 16:3(n-3)); 9,12,15-octadecatetraenoic acid (α-linolenic acid (ALA), 18:3(n-3)); 6,9,12,15-octadecatetraenoic acid (stearidonic acid (STD), 18:4(n-3)); 11,14,17-eicosatrienoic acid (eicosatrienoic acid (ETE), 20:3(n-3)); 8,11,14,17-eicosatetraenoic acid (eicosatetraenoic acid (ETA),20:4 (n-3)); 5,8,11,14,17-eicosapentaenoic acid (eicosapentaenoic acid(EPA), (20:5 (n-3)); 7,10,13,16,19-docosapentaenoic acid(docosapentaenoic acid (DPA), 22:5 (n-3));4,7,10,13,16,19-docosahexaenoic acid (docosahexaenoic acid (DHA), 22:6(n-3)); 9,12,15,18,21-tetracosapentaenoic acid (tetracosapentaenoicacid, 24:5 (n-3)); and 6,9,12,15,18,21-tetracosahexaenoic acid(tetracosahexaenoic acid, 24:6 (n-3)).

Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are found innature in fish oils, and have been used in a variety ofdietary/therapeutic compositions. EPA and DHA are preferred omega-3polyunsaturated fatty acids in the present invention.

Omega-3 Polyunsaturated Fatty Acid Derivatives

As used herein, the term “omega-3 polyunsaturated fatty acidderivative(s)” refers to omega-3 polyunsaturated fatty acids that havebeen reacted with another compound or otherwise modified so that theomega-3 polyunsaturated fatty acid no longer contains a free carboxylicacid. Examples of omega-3 polyunsaturated fatty acid derivatives includesalts, esters (such as alkyl esters including, but not limited to,methyl and ethyl esters) and glycerides of omega-3 polyunsaturated fattyacids. The omega-3 polyunsaturated fatty acid can also be one or more ofthe fatty acid moieties in a phospholipid molecule.

As used herein, the term “glyceride” means a glycerol molecule (i.e.,OHCH₂CHOHCH₂OH) in which one, two or all three of the hydroxyls havebeen esterified with a carboxylic acid, e.g., an omega-3 polyunsaturatedfatty acid. Thus, “triglyceride” refers to glycerides in which all threehydroxyls on the glycerol have been esterified with (the same ordifferent) carboxylic acids. “Diglyceride” refers to glycerides in whichonly two of the hydroxyls on the glycerol have been esterified with (thesame or different) carboxylic acids. “Monoglyceride” refers toglycerides in which only one hydroxyl on the glycerol has beenesterified with a carboxylic acid.

Omega-3 fatty acids are found in nature in the triglyceride form (aglycerol with three fatty acids attached). The natural triglyceride formas found in raw fish oil cannot be readily separated as it occurs intopurified EPA/DHA mixtures by ordinary means such as distillation orcrystallization, because the fatty acids are non-uniformly distributedamong the triglyceride molecules. There are very few, if any, singletriglyceride molecules which are composed of either three EPAs or threeDHAs. Typically, there is a DHA, an EPA, and another fatty acid in atriglyceride molecule. So in order to purify fatty acids to increase theproportion of EPA, DHA, or the total fraction of omega-3's, it isnecessary to hydrolyze the triglycerides to remove at least some fattyacids from the glycerol.

The triglycerides may be converted by any method known to one skilled inthe art without limitation. For example, the triglycerides may beconverted by lipase-catalyzed esterification or lipase catalyzedacidolysis with ethyl or lauryl alcohol, which can selectively leave thehighest amount of EPA and DHA bonded to glycerols and remove othercomponents, leaving EPA and/or DHA as mono- or di-glycerides. The mono-and di-glycerides can then be separated into fractions with differentEPA/DHA ratios, by methods familiar to those skilled in the art such asmultiple stage vacuum distillation and/or fractional crystallization inurea. Advantageously, the purified EPA and DHA esters, afterconcentration, can be reattached to glycerol molecules using enzymaticreacylation to recreate glycerides which are otherwise identical to theoriginal natural triglycerides, except that they are more concentratedin EPA and DHA combined, and they may also have a different ratio ofEPA:DHA than the original fish oil. In some embodiments, at least 60% ofthe omega-3 fatty acids, and preferably 70% or more are converted to thetriglyceride form in the reacylation process. The process may besuccessively repeated with addition of additional catalyst and/or enzymeand additional EPA and DHA until the desired specification proportionsare met. About 60% of triglycerides can be made in the first pass ofreacylation, with most of the remainder of the product being mono- anddi-glycerides.

Polyunsaturated fatty acid triglycerides can be prepared using thefollowing method:

1. Removal of Free Fatty Acids

Raw fish oil in the natural triglyceride molecular form preferably fromanchovies and sardines which contain about 18% EPA and 12% DHA is heatedto 60° C. to decrease viscosity. Sodium oxide is added to bind with freefatty acids in the oil. The mixture is moved to a separator where sodiumoxide bound to free fatty acids (soap) floats to the top and is removed.

The oil is then moved to a second separator where warm water ispreferably added to help remove traces of sodium oxide, as sodium oxidepartitions to water, yet does not interact with the fish oil.

Citric acid may then be added to support splitting the oil from thecombination of water and sodium oxide. The oil is then cooled to 30° C.to protect it from oxidation.

2. Stripping and Purification

Oil is moved to a separate stripping tank, and heated to 200° C. Ethylesters can be added to support the removal of impurities, which bind toethyl esters. Impurities such as dioxins, heavy metals, pcbs, fireretardants, furans and others evaporate and are drawn to the middle ofthe tank where a refrigerating element cools them down and drain them.The added esters are also removed with the impurities.

3. Esterification

The oil is moved to an esterification tank. Ethanol and sodium metal areadded. Sodium metal is a catalyst for breaking off fatty acid strandsfrom the glycerol backbone of the triglyceride fatty acid molecule, thefree fatty acids then combined with ethanol to form ethyl esters. Watercan be added to bind to sodium metal, where the combination of water andsodium metal can be removed.

4. Molecular Distillation

The oil is then moved to a distiller where it is heated to about 120° C.under vacuum. Mono esters and shorter carbon chain molecules move to themiddle where they are cooled and drained, leaving longer carbon chainsremaining as a concentrate. The process typically increases the keyfatty acids by 100% during the first distillation; typically between30-50% during the second distillation. The process can be repeated,although preferably the process is ideally only repeated once, as whenoils undergo heat it can produce oxidation and degradation of the fattyacids in general. Oil waste is also increasing with repeateddistillation, making the process less economical.

5. Reesterification (Reacylation)

The oil is then moved to a reesterification tank where the ethyl estermolecules are reconverted to the triglyceride form, which is the naturalform of that fatty acid molecule. 98% of fats ingested by humans are inthis natural triglyceride form.

The esterification process takes place under low vacuum at about 80° C.Glycerol is added to form the backbone of the glyceride molecules.Nitrogen can be added from the bottom of the tank to cause oil movement.Lipase enzymes are added as catalysts to facilitate the fatty acidsbinding to glycerol. The vacuum in the distillation tank removes theethanol which was previously bound to the fatty acids. The enzymes usedare lipases produced from bacteria or yeast. Perhaps the most effectiveenzymes are Candidan antarctica lipase, and Chromobacterium ViscosumLipase; other enzymes that can be used effectively are Psuedomonas,Mucor miehei, and Candida Cylindracea as well as other enzymes may alsobe used.

The reesterification process typically takes 24 hours, at which pointthe triglycerides typically reaches 60-65%, the remaining glyceridesbeing diglycerides and monoglycerides. Around 3% of the fish oil willremain as ethyl esters, which can be removed together with the ethanol.Adding additional enzymes and/or continuing the enzymatic process canproduce triglyceride molecule concentration of up to 99%. The 60-65%level is probably optimum from an economic point of view.

6. Winterization

The oil in triglyceride form is then moved to a cooling tank at 0° C.,where saturated fats, in particular stearic acid are crystallized. Thepulp is then pumped to a filter press, where the crystals are removed,essentially removing the vast majority of saturated fats from the oil.Depending on the amount of saturated fats in the oil, approximately5-10% of the oil is lost during this process.

7. Bleaching

The oil is then removed to a bleaching tank at 60° C., where bleachingearth or bentonite earth is added to the oil. Any water in the oilevaporates due to the temperature. Any remaining impurities (traceminerals, etc) in the oil attach to the bentonite earth. The oil is thenrun through a bentonite earth filter to remove the bentonite earthtogether with the impurities.

8. Deodorization

Although not a necessary step, it is advantageous to move the oil to adeodorization tank. The tank contains low vacuum at 120° C. Steam isadded at the bottom of the tank, which connects to color and odormolecules (oxidated matter, peroxides) which again travel into thevacuum system and into a residue container. This process gives the oil aneutral color with virtually zero taste and odor.

9. Mixing

The oil is then moved to a separate storage tank. Depending on theconcentration of EPA and DHA desired, various batches can be mixed toyield the concentration desired for the final product.

10. Addition of Antioxidant

Antioxidants, in particular rosemary and mixed tocopherols can be addedto the final oil to dramatically reduce the oxidation process.

11. Drumming

The oil is then drummed in stainless steel drums for storage and toppedoff with nitrogen to remove oxygen and minimize the potential foroxidation.

As used herein, the term “pharmaceutically acceptable” means that thematerial to which it refers is not harmful to the subject.

In some embodiments, the composition of the invention employs a mixtureof omega-3 polyunsaturated fatty acids and/or derivatives that containglycerides. For example, in one embodiment, the mixture contains about35 wt. % triglycerides of EPA and about 25 wt. % triglycerides of DHAand about 10% other omega-3 fatty acids or derivatives thereof. In someembodiments, the mixture contains about 65 wt. % triglycerides of EPA,about 15 wt. % triglycerides of DHA and about 20% other omega-3 fattyacids or derivatives thereof, wherein the EPA and DHA are at least about60% in the triglyceride form and the balance are at least about 90% ofmono- and di-glycerides. In some embodiments, the mixture contains about75% EPA and about 15% DHA, wherein at least about 60% of the combinationof DHA and EPA are in the triglyceride form and the balance is at leastabout 90% mono- and di-glycerides. In another embodiment, the mixturecan contain at least about 60 wt. % of a combination of EPA and DHA in aweight ratio of EPA:DHA of from about 1.4:1 to about 5:1 (for example,2:1 to 5:1, 3:1, 4:1 or 4.3:1) wherein the combination is at least about60% (e.g., at least about 80% or at least about 90% or at least about98%) in the triglyceride form of the fatty acids and the balance is atleast about 80% mono- and di-glycerides. In some embodiments, thecombination is at least about 98% in the triglyceride form, with thebalance being in the monoglyceride and/or diglyceride forms. Some of theabove compositions are disclosed in copending U.S. patent applicationSer. No. 12/015,488, filed Jan. 16, 2008 by Opheim. That patentapplication is incorporated by reference herein in its entirety.

Sources of the omega-3 polyunsaturated fatty acids or derivativesthereof include natural sources including, but not limited to, fish oil(e.g., cod liver oil), flax seed oil, marine oils, sea oils, krill oil,algae and the like. Fish oil is a preferred source.

It is preferred to use a high quality source of omega-3 polyunsaturatedfatty acids or derivatives thereof which is rich in omega-3 oils,preferably containing at least 70% omega-3 oils. The oil can also berich in EPA and DHA. Preferably, at least 75% of the omega oils areEPA+DHA, and more preferably 85% or more are EPA+DHA. The daily dose ofomega-3 oils is about 1 to about 4 grams of omega-3 oil. One possiblesource is a balanced omega-3 formula such as Nordic Naturals, Inc.'sProOmega nutritional supplement, which is 70% omega-3 oils of which50.8% is EPA, 35.1% is DHA and 14.1% is other omega-3 polyunsaturatedfatty acids or derivatives thereof.

One preferred source of omega-3 polyunsaturated fatty acids orderivatives thereof is Pro-EPA nutritional supplement sold by NordicNaturals, Inc. It comprises 69.1% EPA, 16.3% DHA, and 14.6% otheromega-3 polyunsaturated fatty acids or derivatives thereof. Stillanother preferred source of omega-3 polyunsaturated fatty acids orderivatives thereof is Nordic Naturals, Inc.'s Pro-EFA Xtra whichcomprises 56.9% EPA, 14.7% DHA, 17.2% GLA (omega-6 gamma-linolenic acid,i.e., 6,9,12-octadecatrienoic acid (18:3 (n-6) or derivative thereof)),and 11.2% other omega-3 polyunsaturated fatty acids or derivativesthereof. The Pro-EFA Xtra formula adds an omega-6 polyunsaturated fattyacid or derivatives thereof, GLA, and makes a powerful anti-inflammatorymixture.

The Red Yeast Rice extract is water soluble and is not soluble in theomega-3 oil. In some embodiments, a dispersant is used to keep the RedYeast Rice extract in suspension. In some embodiments, the dispersant isabout 70% silica bamboo with lysine made from sunflower oil. A suitablemethod for making a mixture of the present invention is to vigorouslymix 24 weights of Red Yeast Rice extract with 36 weights of fish oil(containing a suitable amount of omega-3 polyunsaturated fatty acids orderivatives thereof), 1.2 weights of bamboo (2%), and 1.8 weights oflysine (3%). The resulting mixture may then be diluted to the desiredomega-3 oil to Red Yeast Rice extract ratio. In some embodiments, theomega-3/Red Yeast Rice extract is then encapsulated in soft gelatincapsules for dispensing. The capsules are typically of such a size thatan integral number of capsules comprise a daily dosage of the mixture.

In some embodiments, a dosage of the omega-3/Red Yeast Rice mixturefurther includes antioxidants such as rosemary, vitamin E, astaxanthine,carnitine, ascorbyl palmitate, tocopherols or other antioxidants knownin the art for stabilizing fish oil and/or omega-3 polyunsaturated fattyacids or derivatives thereof.

Comparison Between Red Yeast Rice Extract and Lovastatin

Red Yeast Rice extract contains monacolin K, the lactone form of thestatin drug Mevacor® (lovastatin). Red Yeast Rice extract has beentested in clinical trials at daily dosages of 1.2 g and 2.4 g. Themonacolin K content of the Red Yeast Rice extract used in the clinicaltrials was 0.20% of the Red Yeast Rice extract. The monacolin K dose wastherefore 2.4 to 4.8 mg/day. At 2.4 mg/day of monacolin K, the totalcholesterol, LDL cholesterol, and triglycerides dropped by 23%, 31%, and34% respectively. At 4.8 mg/day, the reduction was 17%, 23%, and 16%respectively (see Monograph by Thorne Research Inc., AlternativeMedicine Review. Volume 9, Number 1 2004).

Lovastatin has been shown to have a cholesterol lowering effect in dosesranging from 5 to 80 mg/day (see Bates, M, et al, Effectiveness of lowdosage lovastatin in lowering serum cholesterol. Experience with 56patients. Archives of Internal Medicine 1990:150: 1947-1950). A studywas performed to show the effectiveness of low-dose lovastatin inlowering serum cholesterol (see Heber D, et al., Cholesterol loweringeffects of proprietary Chinese red yeast rice dietary supplement, Ann JClin Vutr 1999, 69:231-236). Fifty-six patients were given 20 mg/day oflovastatin for 24 weeks. Total cholesterol fell by 26% and triglyceridesfell by 12%.

Mevacor® (lovastatin) in its package insert reported extensive clinicaltrials at dosages of 10, 20, and 40 mg/day. Total cholesterol wasreduced in the range from 16-24%, LDL was reduced by 21-32%, andtriglycerides were reduced by 10 to 6% (higher reduction observed atlower dosage).

It should be noted that Red Yeast Rice extract at a dosage of 2.4 mg/dayof monacolin K produced better lipid reducing results than Mevacor at10-40 mg/day. It is, therefore, unlikely that the lipid lowering effectswith Red Yeast Rice result from the monacolin K content alone of RedYeast Rice, but are probably attributable in whole or in part to theother monacolins, sterols (beta-sitosterol, campesterol, sigmasterol,and sapogenin), isoflavones, and monounsaturated fatty acids Red YeastRice extract (see Durington P, Bhatnager, et al, An omega-3polyunsaturated fatty acid concentration administered for one yeardecreased triglycerides in simvastin treated patients with CM. Heart2001:85(5) 544-548) This is a particular advantage since the lowerdosage of Red Yeast Rice extract containing HMG-CoA reductase inhibitorcontributes to reduced side effects as well.

Red Yeast Rice extract can be purchased as a nutritional supplement inthe United States. Preferred sources include DRACO Natural Products (539Parrott St., San Jose, Calif. 95112) Red Yeast Rice Extract 10:1, andthe Thorne Research product, Choleast. Purchased Red Yeast Ricepreferably should contain about 0.2 wt. % or more of monacolin K andabout 0.5 wt. % or more of total monacolins.

The compositions of this invention can contain other ingredients besidesthe ingredients recited above. These include, but are not limited to,flavor agents, fillers, surfactants (e.g., polysorbate 80 and sodiumlauryl sulfate), color agents including, e.g., dyes and pigments,sweeteners, antioxidants and additional ingredients.

Flavor Agents

Useful flavor agents include natural and synthetic flavoring sourcesincluding, but not limited to, volatile oils, synthetic flavor oils,flavoring aromatics, oils, liquids, oleoresins and extracts derived fromplants, leaves, flowers, fruits, stems and combinations thereof. Usefulflavor agents include, e.g., citric oils, e.g., lemon, orange, grape,lime and grapefruit, fruit essences including, e.g., apple, pear, peach,banana, grape, berry, strawberry, raspberry, blueberry, blackberry,cherry, plum, pineapple, apricot, and other fruit flavors. Other usefulflavor agents include, e.g., aldehydes and esters (e.g., benzaldehyde(cherry, almond)), citral, i.e., alpha-citral (lemon, lime), neral,i.e., beta-citral (lemon, lime), decanal (orange, lemon), aldehyde C-8(citrus fruits), aldehyde C-9 (citrus fruits), aldehyde C-12 (citrusfruits), tolyl aldehyde (cherry, almond), 2,6-dimethyloctanal (greenfruit), 2-dodecenal (citrus, mandarin) and mixtures thereof, chocolate,cocoa, almond, cashew, macadamia nut, coconut, mint, chili pepper,pepper, cinnamon, vanilla, tooty fruity, mango and green tea. Mixturesof two or more flavor agents may also be employed. When a flavor agentis used, the amount employed will depend upon the particular flavoragent used. However, in general, the flavor agent can constitute fromabout 5% to about 50% by weight of the composition.

Color Agents

Useful color agents include, e.g., food, drug and cosmetic (FD&C) colorsincluding, e.g., dyes, lakes, and certain natural and derived colorants.Useful lakes include dyes absorbed on aluminum hydroxide and othersuitable carriers. Mixtures of color agents may also be employed. When acolor agent is employed, the amount used will depend upon the particularcolor agent used. However, in general, the color agent can constitutefrom about 0.5% to about 5% by weight of the composition.

Sweetening Agent

Natural and/or artificial sweetening agents can also be added to thecomposition. Examples of sweeteners include sugars such as sucrose,glucose, invert sugar, fructose, and mixtures thereof, saccharin and itsvarious salts (e.g., sodium and calcium salt of saccharin), cyclamicacid and its various salts, dipeptide sweeteners (e.g., aspartame),dihydrochalcone, and sugar alcohols including, e.g., sorbitol, sorbitolsyrup, mannitol and xylitol, and combinations thereof. Naturalsweeteners that can be employed include, but are not limited to, luohan, stevia or mixtures thereof. Luo han sweetener is derived from luohan guo fruit (siraitia grosvenorii) that is mainly found in China. Itis about 300 times sweeter by weight than sucrose. Luo han iscommercially available from, e.g., Barrington Nutritionals (Harrison,N.Y.). Stevia is derived from a South American herb, Stevia rebaudiana.It can be up to about 300 times sweeter than sucrose. Because luo hanand stevia have such a sweet taste, only a small amount need be used inthe composition. When a sweetening agent is employed the amount usedwill depend upon the particular sweetening agent used. However, ingeneral, the sweetening agent can constitute from about 0.0005% to about30%, by weight of the composition. When a sweetener having a very sweettaste, such as luo han or stevia, is used, small amounts such as about0.0005% to about 0.1% (for example about 0.005% to about 0.015% or about0.002% to about 0.003%) by weight can be used.

Additional Ingredients

The compositions of the present invention can contain additionalingredients. Examples of such additional ingredients include, but arenot limited to, vitamins, minerals and/or herbs.

As used herein, the term “vitamin” refers to trace organic substancesthat are required in the diet. For the purposes of the presentinvention, the term vitamin(s) include, without limitation, thiamin,riboflavin, nicotinic acid, pantothenic acid, pyridoxine, biotin, folicacid, vitamin B12, lipoic acid, ascorbic acid, vitamin A, vitamin D,vitamin E and vitamin K. Also included within the term vitamin are thecoenzymes thereof. Coenzymes are specific chemical forms of vitamins.Coenzymes include thiamine pyrophosphates (TPP), flavin mononucleotide(FMM), flavin adenine dinucleotive (FAD), Nicotinamide adeninedinucleotide (AND), Nicotinamide adenine dinucleotide phosphate (NADP),Coenzyme A (CoA), Coenzyme Q10 (CoQ10), pyridoxal phosphate, biocytin,tetrahydrofolic acid, coenzyme B12, lipoyllysine, 11-cis-retinal, and1,25-dihydroxycholecalciferol. The term vitamin(s) also includescholine, camitine, and alpha, beta, and gamma carotenes.

As used herein, the term “mineral” refers to inorganic substances,metals, and the like required in the human diet. Thus, the term“mineral” as used herein includes, without limitation, calcium, iron,zinc, selenium, copper, iodine, magnesium, phosphorus, chromium and thelike, and mixtures thereof. Compounds containing these elements are alsoincluded in the term “mineral.”

As used herein, the term “herb” refers to organic substances defined asany of various often aromatic plants used especially in medicine or asseasoning. Thus, the term “herb” as used herein includes, but is notlimited to, black currant, ginsing, ginko bilboa, cinnamon, and thelike, and mixtures thereof.

Other ingredients that can be used include antioxidants, glucosamine andmixtures thereof.

The compositions of this invention are suitable for therapeutic and/ornutritional purposes in treating a subject in need of such treatment. Asused herein, the term “subject” includes, but is not limited to, anon-human animal, such as a cow, monkey, horse, sheep, pig, chicken,turkey, quail, cat, dog, mouse, rat, rabbit, or guinea pig; and a human.

The amount of the composition of the invention that is effective willvary depending upon the condition being treated, and can be determinedby standard clinical techniques. In addition, in vitro or in vivo assayscan optionally be employed to help identify optimal dosage ranges. Theprecise dose to be employed will also depend on the relative amounts ofthe components of the compositions of the invention, route ofadministration, and the seriousness of the condition being treated andshould be decided according to the judgment of the practitioner and eachsubject's circumstances. However, suitable effective dosage amounts forthe compositions of the invention typically are at least about 2 gramsper day, typically administered in the form of capsules containing atleast about 1 gram of the composition per capsule. A typical dose shouldcontain sufficient omega-3 acids or derivatives thereof to provide atleast about 500 mg of a combination of EPA and DHA (or derivativesthereof).

The form in which the composition of the invention is administered tothe subject is not critical. Typically, the composition is administeredas a liquid or in a capsule. Typically, the composition is administeredin the form of individual doses. As used herein, the term “dose”includes both the case where the Red Yeast Rice extract and the omega-3compounds are administered together (such as in the form of a capsulecontaining both components), and the case where the Red Yeast Riceextract and omega-3 compounds are administered separately (but,typically, at essentially the same time). In some embodiments, thecomposition of the invention is administered in the form of a dailydose. However, depending on the severity of the condition being treated,this may not be required, and the period between administration of thedoses may be longer than one day. In addition, the term “administer”includes both the case where a third party administers the dose to thesubject and the case where the subject self-administers the dose.

Examples

In one embodiment a daily dosage is taken to reduce cholesterol andtriglycerides, the dosage comprising 1.2 grams per day of Red Yeast Riceextract and 1700 mg of Nordic Naturals® ProOmega® fish oil supplement(which contains 70% Omega-3 oils which are 85.9% EPA and DHA, so that1700 mg will supply 1000 mg/day of EPA and DHA). Here the weight ratioof fish oil to Red Yeast Rice extract is 1700/1200 (1.4:1). To make themixture add 1200 weights of Red Yeast Rice extract to 1700 weights offish oil. Add 58 weights (2%) of 70% silica bamboo and 87 weights (3%)of lysine and mix thoroughly to make a stable suspension. Fill gelatincapsules with the mixture such that an integral number of capsulesdispenses 3045 grams of the mixture.

In some embodiments, a daily dose is taken, the dose comprising 1.2grams per day of Red Yeast Rice extract and 3400 mg of fish oil such asProOmega® fish oil supplement. Here the weight ratio of fish oil to RedYeast Rice extract is 2.8 and will supply 2000 mg/day of EPA and DHA.

In some embodiments, a daily dose is taken, the dose comprising 2.4grams per day of Red Yeast Rice extract and 3400 mg of fish oil. Here,the weight ratio of fish oil to Red Yeast Rice extract is 1.4.

In some embodiments, a daily dose is taken, the dose comprising 2.4grams per day and 6800 mg of fish oil. Here, the weight ratio of fishoil to Red Yeast Rice extract is 2.8.

Although the present invention has been described in considerable detailwith reference to certain versions thereof, other versions are possible.Therefore the spirit and scope of the appended claims should not belimited to the versions presented herein.

What is claimed is:
 1. A composition comprising (1) EPA, a glyceride ofEPA, or a mixture thereof, and DHA, a glyceride of DHA, or a mixturethereof, wherein at least about 60% w/w of (1) is in reesterifiedtriglyceride form and the weight ratio of EPA in reesterifiedtriglyceride form to DHA in reesterified triglyceride form is from about3:1 to about 5:1; and (2) red yeast rice, wherein the red yeast rice isdispersed in (1) and comprises red yeast rice monacolin compounds,sterols, isoflavones and monounsaturated fatty acids, wherein thecomposition is formulated to provide a daily dose of (1) of from about1000 mg to about 4000 mg and a daily dose of (2) of from about 1200 mgto about 2400 mg.
 2. The composition of claim 1, wherein the red yeastrice comprises monacolin K and at least one other red yeast ricemonacolin compound.
 3. The composition of claim 2, wherein the red yeastrice comprises at least about 0.1 wt % monacolin K.
 4. The compositionof claim 2, wherein the red yeast rice comprises at least about 0.2 wt %monacolin K.
 5. The composition of claim 2, wherein the red yeast ricecomprises at least about 0.4 wt % monacolin K.
 6. The composition ofclaim 1, wherein the red yeast rice comprises all of the monacolincompounds in red yeast rice.
 7. The composition of claim 6, wherein thered yeast rice comprises at least about 0.4 wt % total red yeast ricemonocolins.
 8. The composition of claim 1, wherein at least about 60% of(1) is in the reesterified triglyceride form and the balance is mono-and diglycerides.
 9. The composition of claim 1, wherein at least about80% of (1) is in the reesterified triglyceride form.
 10. The compositionof claim 1, wherein at least about 90% of (1) is in the reesterifiedtriglyceride form.
 11. The composition of claim 1, wherein at leastabout 98% of (1) is in the reesterified triglyceride form.
 12. Thecomposition of claim 1, further comprising a dispersing agent.
 13. Thecomposition of claim 1, formulated into a gelatin capsule, wherein thedaily doses of (1) and (2) are provided in an integral number ofcapsules.
 14. The composition of claim 1, further comprising anantioxidant.
 15. The composition of claim 1, further comprising otheromega-3 polyunsaturated fatty acids or derivatives thereof.
 16. Thecomposition of claim 1, wherein the weight ratio of EPA in reesterifiedtriglyceride form to DHA in reesterified triglyceride form is about 4:1.17. The composition of claim 1, wherein, in the component (1) inreesterified triglyceride form, three fatty acid molecules, eachindependently selected from the group consisting of EPA and DHAmoieties, are attached to a glycerol backbone.
 18. The composition ofclaim 1, wherein the reesterified triglyceride form of component (1) isprepared by a process comprising removing fatty acids from a glycerolbackbone of a triglyceride and then reesterifying EPA and/or DHAmoieties onto the glycerol backbone.
 19. The composition of claim 14,wherein the antioxidant is selected from one or more of rosemary,vitamin E, astaxanthine, carnitine, ascorbyl palmitate, and tocopherols.20. The composition of claim 1, wherein the composition is formulated toprovide a daily dose of (1) of about 1000 mg and a daily dose of (2) ofabout 1200 mg.
 21. The composition of claim 1, wherein the compositionis formulated to provide a daily dose of (1) of about 2000 mg and adaily dose of (2) of about 1200 mg.
 22. The composition of claim 1,wherein the composition is formulated to provide a daily dose of (1) ofabout 4000 mg and a daily dose of (2) of about 2400 mg.
 23. A method ofreducing serum cholesterol, triglycerides, or both, comprising orallyadministering the composition of claim 1 to a human subject in needthereof in an amount effective to provide a daily dose of (1) of fromabout 1000 mg to about 4000 mg and a daily dose of (2) of from about1200 mg to about 2400 mg.
 24. The method of claim 23, wherein thecomposition is administered in an amount effective to provide a dailydose of (1) of about 1000 mg and a daily dose of (2) of about 1200 mg.25. The method of claim 23, wherein the composition is administered inan amount effective to provide a daily dose of (1) of about 2000 mg anda daily dose of (2) of about 1200 mg.
 26. The method of claim 23,wherein the composition is administered in an amount effective toprovide a daily dose of (1) of about 4000 mg and a daily dose of (2) ofabout 2400 mg.